Vaccine prepared utilizing human parainfluenza virus type 2 vector
AssigneesBiocomo Inc., Mie University
InventorsMasayuki Fukumura, Junpei Ohtsuka, Tetsuya Nosaka, Masato Tsurudome, Mitsuo Kawano, Ken-ichiro Hara
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Abstract
Disclosed are: a virus vector in which a gene encoding an antigenic polypeptide is integrated in human parainfluenza virus type 2 gene, wherein the antigenic polypeptide is expressed in the form of a fusion protein with a viral structural protein; and a method for producing the same. The virus vector of the present invention contains a quantitatively large amount of the antigenic peptide on the virus particle and can efficiently deliver the antigenic polypeptide to a target cell.
Claims16
A virus vector in which a gene encoding an antigenic polypeptide is integrated in a gene of an F protein-defective human parainfluenza virus type 2, wherein the antigenic polypeptide is expressed in the form of a fusion protein with a viral structural protein or a portion thereof, and the virus vector requires that F protein be supplied in trans by a packaging cell in order to produce virus particles having infectious ability, wherein the F protein-defective virus lacks F gene or an extramembrane domain of the F gene.
A method for producing a virus vector in which a gene encoding an antigenic polypeptide is integrated in a gene of an F protein-defective human parainfluenza virus type 2, wherein the antigenic polypeptide is expressed in the form of a fusion protein with a viral structural protein or a portion thereof, and the virus vector requires that F protein be supplied in trans by a packaging cell in order to produce virus particles having infectious ability, wherein the F protein-defective virus lacks F gene or an extramembrane domain of the F gene, the method comprising the steps of:
- (1) coculturing:
- (i) a Vero cell transfected with (a) an F gene of human parainfluenza virus type 2 or (b) an F gene of human parainfluenza virus type 2 fused with a gene encoding an antigen, wherein the Vero cell stably expresses the F gene or the F gene fused with the gene encoding the antigen;
- and
- (ii) an F gene-defective human parainfluenza virus type 2 in which a gene encoding the fusion protein is integrated in a viral gene;
- and
- (2) isolating virus particles from the culture supernatant;
- wherein the fusion protein is a fusion protein with one or more selected from HN protein, F protein, and M protein of the virus, or a portion thereof.
A method for producing a virus vector in which a gene encoding an antigenic polypeptide is integrated in a gene of an F protein-defective human parainfluenza virus type 2, wherein the antigenic polypeptide is expressed in the form of a fusion protein with a viral structural protein or a portion thereof, and the virus vector requires that F protein be supplied in trans by a packaging cell in order to produce virus particles having infectious ability, wherein the F protein-defective virus lacks F gene or an extramembrane domain of the F gene, the method comprising the steps of:
- (1) coculturing:
- (i) a Vero cell transfected with (a) an F gene of human parainfluenza virus type 2 or (b) an F gene of human parainfluenza virus type 2 fused with a gene encoding an antigen, wherein the Vero cell stably expresses the F gene or the F gene fused with the gene encoding the antigen;
- and
- (ii) an F gene-defective human parainfluenza virus type 2 in which a gene encoding the fusion protein is integrated in a viral gene;
- and
- (2) isolating virus particles from the culture supernatant;
- wherein the fusion protein is a fusion protein with an intracellular domain of the HN protein or the F protein of the virus.
A method for producing a virus vector in which a gene encoding an antigenic polypeptide is integrated in a gene of an F protein-defective human parainfluenza virus type 2, wherein the antigenic polypeptide is expressed in the form of a fusion protein with a viral structural protein or a portion thereof, and the virus vector requires that F protein be supplied in trans by a packaging cell in order to produce virus particles having infectious ability, wherein the F protein-defective virus lacks F gene or an extramembrane domain of the F gene, the method comprising the steps of:
- (1) coculturing:
- (i) a Vero cell transfected with (a) an F gene of human parainfluenza virus type 2 or (b) an F gene of human parainfluenza virus type 2 fused with a gene encoding an antigen, wherein the Vero cell stably expresses the F gene or the F gene fused with the gene encoding the antigen;
- and
- (ii) an F gene-defective human parainfluenza virus type 2 in which a gene encoding the fusion protein is integrated in a viral gene;
- and
- (2) isolating virus particles from the culture supernatant;
- wherein the antigenic polypeptide is
- 1) fused on the C-terminal side of the HN protein and expressed outside the vector envelope,
- 2) fused on the C- or N-terminal side of the F protein and expressed inside the vector envelope, or
- 3) fused on each of the C-terminal side of the HN protein and the C- or N-terminal side of the F protein or the N-terminal side of the HN protein and expressed both outside and inside the vector envelope.
A method for producing a virus vector in which a gene encoding an antigenic polypeptide is integrated in a gene of an F protein-defective human parainfluenza virus type 2, wherein the antigenic polypeptide is expressed in the form of a fusion protein with a viral structural protein or a portion thereof, and the virus vector requires that F protein be supplied in trans by a packaging cell in order to produce virus particles having infectious ability, wherein the F protein-defective virus lacks F gene or an extramembrane domain of the F gene, the method comprising the steps of:
- (1) coculturing:
- (i) a Vero cell transfected with (a) an F gene of human parainfluenza virus type 2 or (b) an F gene of human parainfluenza virus type 2 fused with a gene encoding an antigen, wherein the Vero cell stably expresses the F gene or the F gene fused with the gene encoding the antigen;
- and
- (ii) an F gene-defective human parainfluenza virus type 2 in which a gene encoding the fusion protein is integrated in a viral gene;
- and
- (2) isolating virus particles from the culture supernatant;
- wherein the antigenic polypeptide is fused with a membrane or intracellular domain of the HN protein or the F protein of the virus so that the antigenic polypeptide is included on the vector envelope.
A method for producing a virus vector in which a gene encoding an antigenic polypeptide is integrated in a gene of an F protein-defective human parainfluenza virus type 2, wherein the antigenic polypeptide is expressed in the form of a fusion protein with a viral structural protein or a portion thereof, and the virus vector requires that F protein be supplied in trans by a packaging cell in order to produce virus particles having infectious ability, wherein the F protein-defective virus lacks F gene or an extramembrane domain of the F gene, the method comprising the steps of:
- (1) coculturing:
- (i) a Vero cell transfected with (a) an F gene of human parainfluenza virus type 2 or (b) an F gene of human parainfluenza virus type 2 fused with a gene encoding an antigen, wherein the Vero cell stably expresses the F gene or the F gene fused with the gene encoding the antigen;
- and
- (ii) an F gene-defective human parainfluenza virus type 2 in which a gene encoding the fusion protein is integrated in a viral gene;
- and
- (2) isolating virus particles from the culture supernatant;
- further comprising the step of subjecting the virus to nucleic acid inactivation treatment.
Dependencies are inferred from the claim text.